Infant nutrition with hydrolysed protein, ionic calcium and palmitic acid

ABSTRACT

The present invention relates to a composition for use in formulae for infants or young children comprising partially hydrolysed protein, ionic calcium and an oil mix comprising palmitic acid esterified to triacylglycerols wherein the palmitic acids esterified in the sn-2 position of triacylglycerols in an amount of at least 20% by weight of total palmitic acid. Furthermore, the invention relates to an infant formula comprising said composition and the use of said composition or infant formula.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a composition for use in formulaes forinfants or young children comprising partially hydrolysed protein, ioniccalcium and an oil mix comprising palmitic acid esterified totriacylglycerols, wherein the palmitic acid comprises palmitic acidesterified in the sn-2 position of triacylglycerols in an amount of atleast 20% by weight of total palmitic acid. Furthermore, the inventionrelates to an infant formula comprising said composition and the use ofsaid composition or infant formula.

BACKGROUND OF THE INVENTION

Mother's milk is recommended for all infants as a sole source ofnutrition for up to the age of 4-6 months of age, since human breastmilk supplies vital nutrients to infants. However, in some casesbreastfeeding is inadequate or unsuccessful or inadvisable for medicalreasons, or the mother chooses not to breast feed either at all or for aperiod of more than a few weeks. Infant formulations have been developedfor these situations.

Research into the components of human milk has been going on for manyyears and it is by no means complete even now. The research howevershows that there are differences between the nutrients present in humanmilk and the ones used in the commercially available infant formulas onthe market today. For example, the type of lipid composition can differsignificantly. The lipids present in human breast milk contains forexample a large amount of palmitic acid esterified on the sn-2 (beta)position of the triacylglycerol, i.e. 68-72% of palmitic acid in humanmilk is esterified in the sn-2 position. In infant formulas, vegetableoils are typically used as the lipid source. The source of palmitic acidin the infant formulas used today comprises a high amount (i.e. 80%) ofpalmitic acid esterified in the sn-1(3) (alpha) position of thetriacylglycerol.

This difference in oils and fats and in particular the difference inregiospecific distribution of palmitic acid causes a difference in thedigestion of fats in infants. Infants fed with infant formulationscomprising an oil mix comprising palmitic acid having a high amount ofsaid palmitic acid in the sn-1(3) position of the triacylglycerol havebeen shown to lead to an increased calcium excretion in the faeces ascompared to infants fed with a human breast milk. Palmitic acid in thesn-1 and sn-3 position are most likely to be cleaved and form free fattyacids. Free palmitic acid can bind calcium and form insoluble palmititicacid soaps in the intestine. Excretion of these soaps in the feces maybe partially responsible for harder stools experienced by some formulafed infants. Harder stools are a component of functional constipation ininfants and young children.

Further, calcium loss in a population such as infants may increase therisk of insufficient bone mineralization.

Some studies are known where infants fed with infant formulas comprisinga high relative proportion of sn-2 palmitate, such as more than 66% sn-2palmitate of total amount of palmitic acid (and lower than 34% sn-1(3)palmitate) were observed to have a significant reduction of calciumexcreted in the faeces as compared to the calcium extreted in infantsfed woth a standard formula. On the contrary, infants fed with an infantformula having a more moderate content of sn-2 palmitate, approximately44% of total palmitic acid, showed no significant decrease in faecalcalcium as compared to infants fed with a standard formula.

In Carnielli et al “Feeding premature newborn infants with palmitic acidin amounts and stereoisomeric position similar to that of human milk;effects on fat and mineral balance”, Am J Clin Nutr. May 1995 vol. 61no. 5, page 1037-1042 is disclosed that an infant formula comprisingsn-2 palmitates in an amount close to that in human breast milk resultedin a lower calcium excretion as compared to when infants were fed withan infant formula having a high amount of sn-1(3) palmitate.

In Carnielli et al “Structural position and amount of palmitic acid ininfant formulas: effects on fat, fatty acid, and mineral balance” J.Pediatr Gastroenterol Nitr. 1996, December; 23(5):553-60, is disclosed astudy showing that infants fed with an infant formula comprising 24%palmitic acid having 66% esterified in the sn-2 position oftriacylglycerol, would have a faecal calcium excretion significantlylower than for infants fed with an infant formula comprisingintermediate and low amounts of sn-2 palmitate out of total palmiticacid (24% palmitic acid and 39% esterified in the sn-2 position oftriacylglycerol) and (20% palmitic acid having 13% esterified in thesn-2 position of the triacylglycerol).

Furthermore, studies have shown that infant formulations comprising anamount of 36% of total palmitic acid esterified in the sn-2 position oftriacylglycerols and lower and thus 64% of palmitic acid esterified inthe sn-1(3) position of triacylglycerold and higher, will result in nosignificant decrease in the faecal calcium excretion.

Thus, the palmitic acid as well as its regiospecific distribution (sn-2versus sn-1(3)) is important for reduction of calcium excreted infaeces.

However, there is a need in the art for compositions for use informulaes for infants or young children where the amounts of calciumexcreted in stools are even lower and more comparable with the calciumlevels excreted by infants fed with human milk.

SUMMARY OF THE INVENTION

Thus, an object of the present invention is to provide a composition foruse in formulaes for infants or young children, and in human milkfortifier, which can result in similar low amount of calcium excreted infaeces as when an infant is fed with human breast milk.

In particular, it is an object of the present invention to provide acomposition that solves the above mentioned problems of the prior artwith calcium excretion and which composition further decrease the amountof calcium excreted by an infant fed with an infant formula.

Thus, one aspect of the invention relates to a composition for use informulaes for infants or young children comprising partially hydrolysedprotein, ionic calcium and an oil mix comprising palmitic acidesterified to triacylglycerol, wherein the palmitic acid comprisespalmitic acid esterified in the sn-2 position of triacylglycerol in anamount of at least 20% by weight.

Another aspect of the present invention is to provide an infant formulacomprising the composition according to the invention.

Yet another aspect of the present invention relates to the compositionor infant formula according to the invention for use in administrationto an infant or young child to reduce calcium excretion by said infantor young child.

Still another aspect of the present invention relates to the compositionor infant formula according to the invention for use in administrationto an infant or young child to increase calcium homeostasis, increasecalcium absorption, increase calcium retention, increase calciumutilization and/or reduce the formation of palmitic acid soaps.

In still another aspect, the present invention relates to thecomposition or infant formula according to the invention for use inadministration to an infant or young child to soften stools, preventand/or reduce the risk of hard stools, prevent and/or reduce the risk ofconstipation, improve feeding tolerance, decrease the frequency andduration of crying and fussing and alleviate digestive discomfort andcolic.

In still another aspect, the present invention relates to thecomposition or infant formula according to the invention for use inadministration to an infant or young child to improve sleep duration,improve sleep quality and quantity, improve infant and parental qualityof life and reduce maternal anxiety.

In a further aspect, the present invention relates to the composition orinfant formula according to the invention for use in administration toan infant or young child to increase bone mineralization, increase bonestrength, and/or increase bone mineral density.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Prior to discussing the present invention in further details, thefollowing terms and conventions will first be defined:

Numerical ranges as used herein are intended to include every number andsubset of numbers contained within that range, whether specificallydisclosed or not. Further, these numerical ranges should be construed asproviding support for a claim directed to any number or subset ofnumbers in that range. For example, a disclosure of from 1 to 10 shouldbe construed as supporting a range of from 1 to 8, from 3 to 7, from 1to 9, from 3.6 to 4.6, from 3.5 to 9.9 and so forth. All references tosingular characteristics or limitations of the present invention shallinclude the corresponding plural characteristic or limitation, and viceversa, unless otherwise specified or clearly implied to the contrary bythe context in which the reference is made.

In the context of the present invention, the term “ratio” by weight(weight/weight) refers to the ratio between the weights of the mentionedcompounds. For example, a mixture comprising 60 g whey and 40 g caseinwould have a weight ratio which is equal to 60:40, which is equal to 3:2or 1.5 (that is 3 divided with 2). Similarly, a mixture of 50 g whey and50 g casein would have a ratio by weight of whey and casein of 50:50,which is equal to 1:1 or 1 (that is 1 divided with 1).

The term “and/or” used in the context of the “X and/or Y” should beinterpreted as “X”, or “Y”, or “X or Y”.

As used in this specification, the words “comprising”, “comprises” andsimilar words are not to be interpreted in an exclusive or exhaustivesense. In other words they are intended to mean “including, but notlimited to”.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art.

The term “infant” will in the context of the present invention mean achild under the age of 12 months.

The term “young child” refers to a child in the age from 12 months to 4years.

In the context of the present invention, the infant may be any terminfant or preterm infant. In an embodiment of the invention, the infantis selected from the group of preterm infants and term infants.

The term “infant formula” as used in the context of the presentinvention refers to a nutritional composition intended for infantsduring the forst months of life and as defined in Codex Alimentarius,(Codex STAN 72-1981) and Infant Specialities (incl. Food for SpecialMedical purposes) as defined in Codex Alimentarius, (Codex STAN72-1981).

The term “follow-on formula” are given to formulas designed to be usedfrom the age of 6 to 12 months of age. The follow-on formula istypically given in combination with the infant is given other types offood, such as vegetable and/or fruit puré.

The term “growing-up milk” is given to formulas designed to be used fromthe age of one year onwards. It is generally a milk-based beverageadapted for the specific nutritional needs of young children.

The term “composition for use in formulaes for infants or youngchildren” refers in the context of the present invention to either aformula as such, i.e an infant formula, which comprises all nutrientsnecessary in order to meet the standards of being an infant formula asdefined in the Codex Alimentarius. Further, the “composition for use informulaes for infants or young children” may be a composition comprisingnutrients, which together with other compositions of nutrients can bemixed to prepare a formula, i.e. such “composition for use in infantformulation” can be added to a mixture, which is intended to be used asan infant formula. Further, the “composition for use in formulaes” canbe a supplement or a fortifier to an infant formula, follow-on formula,growing-up milk or human milk.

Oil Mix:

In the context of the present invention, the term “oil mix” refers to amixture of vegetable fats and oils, dairy fats and/or animal fatscomprising triacylglycerols as the main source of lipids. The oil mixmay be a mixture of one or more vegetable oils and fats, for exampleselected from the group consisting of palm oil, palm kernel oil,esterified vegetable oils, such as interesterified palm oil, animal fatand animal fat fractions. The oil mix may also comprise other vegetableoils and fats suitable for infant nutrition. For example the oil mix maycomprise one or more selected from the group of soyabean oil, coconutoil, rapeseed oil, canola oil, sunflower oil, safflower oil and milkfat.

In an aspect of the present invention, the oil mix used comprisespalmitic acid, wherein the proportion of palmitic acid esterified in thesn-2 position of triacylglycerol is least 20% by weight of totalpalmitic acid.

Palmitic Acid:

In the context of the present invention, the term “palmitic acid” refersto the saturated fatty acid, 16:0. The term “palmitate” may also be usedfor “palmitic acid” when esterified to glycerol such as intriacylglycerols. Most palmitic acid in lipid compositions esterifed totriacylglycerols and may be positioned on either the external (sn-1(3))or internal (sn-2) position of the triacylglycerols. When palmitic acidis esterified in the (external) first or third position of atriacylglycerol, the palmitic acid is in the sn-1(3) position of thetriacylglycerol. Such palmitic acid is in the context of the presentinvention referred to as sn-1(3) palmitate. Palmitic acid esterified inthe internal position of a triacylglycerol is a palmitic acid with theposition of palmitic acid in the sn-2 position of the tricylglycerol,and such palmitic acid is in the context of the present inventionreferred to as sn-2 palmitate. The oil mix according to the presentinvention comprises triacylglycerols and some of the triacylglycerolshas palmitic acids esterified.

In the context of the present invention, the term “amount sn-2 palmiticacid” refers to the amount of beta palmitic acid based on total amountof palmitic acid. Similarly, the term “amount sn-1(3) palmitic acid”refers to the amount of alpha palmitic acid based on total amountpalmitic acid.

In an embodiment according to the present invention, the fat blendcomprises palmitic acid in an amount of at least 8% by weight of totalamount of fatty acids, such as at least 10% by weight, preferably atleast 12% by weight, such as at least 15% by weight, even morepreferably at least 18% by weight. [Jonathan will get back with theminimum amount] In a further embodiment according to the presentinvention, the oil mix comprises palmitic acid in an amount of from 8%to 30% by weight, such as in an amount of from 10 to 26% palmitic acidby weight, preferably in an amount of from 12 to 25% by weight. Inparticular, the oil mix comprises from 20 to 24% by weight of palmiticacid.

In human milk, more than 98% of fat is in form of triacylglycerols whichcontain saturated and unsaturated fatty acids. The main saturated fattyacid in human milk is palmitic acid which account for 22-25% of thetotal amount of fatty acids.

In human milk, about 68-72% of the palmitic acid is esterifed in sn-2position of triacylglycerol.

In palm olein, which is a palm oil fraction widely used in infantformulation comprising a high amount of palmitic acid, the amount ofsn-1(3) palmitate is 90-95% and thus only about 5-10% of palmitic acidis esterified in the sn-2 position of triacylglycerols.

In most commercially available infant formulas, the major part, about80-90% of the palmitic acid present, is esterified in the sn-1(3)position of triacylglycerols, i.e. is present as sn-1(3) palmitate.

However, it has been found out that the position of palmitic acid intriacylglycerols has an influence on the absorption of minerals, such asfor example absorption of calcium, because free palmitic acid formscomplexes with minerals. When palmitic acid is esterified in the sn-1(3)position of triacylglycerols, free palmitic acid during digestion isreleased from the triacylglycerol and forms insoluble complexes withminerals such as calcium if sufficiently present in the diet (this iscalled palmitic acid soaps). These palimitic acid soaps are notabsorbable and are therefore lost in the faeces. The loss of palmiticacid soaps leads to mineral loss as well as poor nutrient absorption.

Palmitic acid soaps may be expressed in many ways, for example ascalcium soaps, palmitate soaps, calcium-fatty acid complexes orinsoluble complexes. These terms may be used interchangeable, but shouldbe understood as the same.

On the contrary, when palmitic acid is esterified in the sn-2 positionof the triacylglycerols, unsaturated fatty acids esterified in thesn-1(3) position of the triacyglycerols are released during digestionand well-absorbed, therefore avoiding palmitic acid soap formation.Furthermore, sn-2 palmitate is well-absorbed as monoacylglycerol. Hence,if a large amount of palmitic acid is in the sn-1(3) position oftriacylglycerol, a large amount of calcium will be excreted in thefaeces. Studies has shown that using an oil mix comprising palmitic acidhaving an amount of sn-2 palmitate in a high amount, such as in anamount similar to the content in human milk, preferably above 65-70%will result in an decrease in calcium excretion as compared to using oilmixes having lower amounts of sn-2 palmitate.

A high amount of sn-2 palmitate is considered to be an amount of sn-2palmitate close to the amount present in human milk, i.e. a high amountof sn-2 palmitate is considered to be in the range of 68-80% by weightof total palmitic acid, such as at least above 65% sn-2 palmitate oftotal palmitic acid.

The inventors of the present invention have surprisingly found out thata combination of an oil mix with palmitic acids, hydrolysed proteins andionic calcium have a synergistic effect on absorption of calcium whencombined with sn-2 palmitate lipids. The inventors have found out thatwhen an oil mix comprising sn-2 palmitates in both moderate amounts,such as about 20-60% by weight, and in higher amounts (65-80%), aredigested by an infant or young child in combination with partiallyhydrolysed protein and ionic calcium, the amounts of calcium excreted instools are close to the amount excreted by infants fed with human milk.

Thus, one aspect of the present invention is to provide a compositionfor use in formulaes for infants or young children comprising partiallyhydrolysed protein, ionic calcium and a oil mix comprising palmitic acidesterified to triacylglycerols, wherein the palmitic acid comprisespalmitic acid esterified in the sn-2 position of palmitic acid in anamount of at least 20% by weight of total palmitic acid.

In an embodiment according to the invention, the oil mix comprises sn-2palmitate in an amount of 2-15% of total fatty acids present in the oilmix and sn-1(3) palmitate in an amount of 5-14% of total fatty acids inthe oil mix.

In a furhter embodiment of the invention, the oil mix comprises palmiticacid, wherein the amount of palmitic acid comprises from 20% to 80% byweight of sn-2 palmitate. In a preferred embodiment, the palmitic acidcomprises sn-2 palmitate in an amount of 25% to 75% by weight, such assn-2 palmitate in an amount of 30% to 70% by weight, preferably sn-2palmitate in an amount of 35% to 65% by weight.

An oil mix comprising the specific amount of sn-2 palmitate may beprepared by mixing different vegetable oils or by mixing vegetable oilswith milk fat. An example of an oil mix could be a mix of cow's milk fatand one or more of vegetable oils, such as sn-2 palmitate, palm oil,coconut oil, soybean oil, high oleic safflower oil and sunflower oil.These fats and oils may be blended in ratios sufficient to provide asimilar fatty acid profile to that found in human milk.

A commercially available composition sold by Lipid Nutrition is Betapol™B-55, which is a triglyceride mixture derived from vegetable oil inwhich at least 54% of the palmitic acid is in the sn-2 position of theglycerol molecule. In one embodiment, the oil mix present in thecomposition according to the present invention is a mix of Betapol™ B-55and other vegetable oils. Those skilled in the art will know that thepercentage of the sn-2 palmitic acid used and the total amount ofpalmitate in the formula may vary, and that a different sn-2 palmitateoil may be used, without departing form the spirit and scope of theinvention.

In another embodiment of the invention, the oil mix furthermorecomprises stearic acid, i.e. triacylglycerols having stearic acidattached thereto through ester bonds.

Protein:

In an aspect of the invention, the composition comprises a partiallyhydrolysed protein.

The inventors of the present invention have surprisingly found out thathydrolysed proteins have a positive influence on the calcium absorptionand that hydrolysed proteins in combination with palmitic acids candecrease the palmitic acid soap formation and thus decrease the calciumexcretion even though the palmitic acids is not all in the sn-2position. Further, the inventors have found out that ionic calcium incombination with palmitic acid and hydrolysed proteins further minimizesthe calcium excretion. It has surprisingly been found that infants fedwith a formula comprising partially hydrolysed protein, ionic calciumand triacylglycerols with palmitic acid results in having an excretionof calcium which is lower than infants fed with a high amount of sn-2palmitate and intact or hydrolysed protein, but are more similar tocalcium excreted by infants fed with human milk.

Without being bound by any theory, the inventors of the presentinvention believes that peptides resulting from hydrolysing a proteinmay have a less protective effect against proteolysis of digestiveenzymes as compared to intact protein. Thus, peptides from proteinhydrolysates may not protect the digestive enzyme lipase fromproteolysis during digestion while intact proteins will protetectlipase. Thus, hydrolysed protein peptides in combination with an oil mixcomprising palmitic acid will result in in less release of sn-1(3)palmitic acid by lipase, and thus prevent soap formation and improvecalcium absorption.

This effect of hydrolysed proteins on the digestive enzymes are furtherimproved by addition of ionic calcium. Without being bound by anytheory, the present inventors believe that ionic calcium form bonds topeptides from hydrolysis of protein. These calcium bound peptides willresult in an even poorer protection of lipase, and thus an even furtherimpairing of lipase activity, than made by unbound peptides alone fromhydrolysed proteins. Thus, when lipase is less protected, less palmiticacids are released from the sn-1 and sn-3 position of the triglycerideand less calcium soaps are formed. The palmitic acids will maintainbound to the triglyceride and absorbed. As a consequence, more calciumis excreted in the faeces.

In the context of the present invention, the term “protein” refers toone or more proteins.

The protein used in the present invention may be any protein suitablefor being administered to infants. The source of protein may for examplebe one or more selected from the group of whey protein, casein, soybeanprotein, pea protein, rice protein or protein from legumes. However, itis preferred that the protein source is based on cow's milkproteins suchas whey, casein or mixtures thereof.

In an embodiment of the present invention, the protein is a dairyprotein and in a preferred embodiment, the protein is a combination ofcasein and whey protein.

Casein is a dairy protein commonly found in mammalian milk. Casein makesup about 80% of the proteins in cow's milk and about 40% of the proteinsin human milk.

Whey protein is a mixture of globular proteins isolated from whey, whichis a liquid material made as a by-product under cheese making. Wheyproteins makes up about 20% of the proteins in cow's milk, where about60% of the proteins in human milk is whey proteins.

Whey proteins is typically a mixture of many proteins, for examplebeta-lactoglobulin, alpha-lactalbumin, lactoferrin and bovine serumalbumin and immunoglobulins.

In another preferred embodiment of the invention, the protein is wheyprotein.

The whey protein may be in the form of for example a whey proteinisolate (WPI) or a whey protein concentrate (WPC).

In an aspect of the present invention, the protein is partiallyhydrolysed protein.

In an embodiment of the present invention, the partially hydrolysedprotein is partially hydrolysed whey protein.

The term “hydrolysed protein” refers to protein that has been subjectedto hydrolysis to break down the protein into a mixture of peptides andfree amino acids. Hydrolysis of proteins is commonly know and can bedone by many different methods, for example by enzymatic hydrolysis byusing an enzyme such as pancreatic protease to stimulate the naturallyoccurring hydrolytic process. Hydrolysis may also be by prolongedboiling in a strong acid or a strong base. In the context of the presentinvention, the partially hydrolysed protein may be produced by anysuitable method known in the art. For example, a whey proteinhydrolysate may be prepared by enzymatically hydrolyse the whey fractionin two steps as described in EP 0322589 A1. For extensively hydrolysingprotein, the whey protein may be subjected to triple hydrolysis usingfirst the enzyme Alcalase 2.4 L (EC 940459), then Neutrase 0.5 L andthen pancreatic protease at temperatures about 55° C.

An example of a whey protein hydrolysate which could be used in thepresent invention is a whey protein hydrolysate comprising about 79%protein and has a degree of hydrolysation being about 13%

In the context of the present invention “partially hydrolysed” refers toproteins, which has been subjected to hydrolysis, where only a portionof the peptide bonds in the protein has been hydrolysed. The partiallyhydrolysis of protein is measured as the degree of hydrolysis.

The term “degree of hydrolysis” of a protein refers to the number ofpeptide bonds in the intact protein which are cleaved during thehydrolysis divided by the number of peptide bonds in the intact proteinexpressed as a percentage.

The term “intact” means in the context of the present invention,proteins where the molecular structure of the protein(s) is not alteredaccording to the conventional meaning of intact proteins. By the term“intact” is meant that the main part of the proteins are intact, i.e.the molecular structure is not altered, for example at least 95% of theproteins are not altered, such as at least 98% of the proteins are notaltered.

In an embodiment of the invention the partially hydrolysed protein has adegree of hydrolysation from 8 to 24%, preferably from 11 to 18%, suchas from 13 to 16%.

Partially hydrolysed protein comprises protein where essentially allprotein has been hydrolysed to some degree. However, the partiallyhydrolysed protein may comprise some intact protein, where the peptidebonds in some proteins under hydrolysis is broken. In the context of thepresent invention, a partially hydrolysed protein hydrolysate comprisesat most 10% intact protein, such as at most 5% intact protein,preferably at most 3% intact protein, even more preferably at most 1%intact protein. The term “essentially all” refers to that the main parthas been hydrolysed, such as at least 95% of the protein has beenhydrolysed preferably at least 98% of the protein has been hydrolysed.

In an embodiment of the invention, the composition, for example aninfant formula, may comprise both a partially hydrolysed proteinhydrolysate and another protein source which comprises intact protein.For example, the source of protein may comprise both intact protein andpartially hydrolysed proteins, such as a for example a whey proteinisolate. Further, the composition or infant formula according to thepresent invention may comprise a partially hydrolysed proteinhydrolysate and another protein source, said other protein sourcecomprising intact protein. For example partially hydrolysed whey proteinisolate and skim milk protein.

The amount of hydrolysed protein should be at least 50% based on totalprotein, such as at least 60%, preferably at least 70%, such as at least80%, even more preferably at least 90%.

Skim milk (may also be called skimmed milk) is a product made from wholemilk where essentially all milk fats are removed from the product. Inskim milk the amount of fat is from zero to 0.3%. Thus, skim milk willcomprise both whey protein and casein.

Another example can be a combination if partially hydrolysed wheyprotein isolate and whole milk protein.

In an embodiment of the invention, the partially hydrolysed protein ispresent in an amount of at least 5 g per litre, such as least 7 g perlitre, preferably at least 9 g per litre. For example, the protein ispresent in the composition in an amount of 5 g to 35 g per litre, suchas from 8 g to 30 g per litre, preferably 9 to 17 g per litre, or 15 to30 g per litre.

In a further embodiment of the present invention, the ratio between thepartially hydrolysed protein and sn-2 palmitate is from 1.5:1 to 35:1 byweight, such as from 2.0:1 to 30:1, preferably from 2.5:1 to 25:1, evenmore preferably from 2.5:1 to 15:1, such as from 2.5:1 to 10:1 byweight.

In a further embodiment of the present invention, the ratio between thepartially hydrolysed protein and sn-1(3) palmitate is from 2:1 to 20:1by weight, such as from 2.5:1 to 15:1, preferably from 3:1 to 10:1 byweight.

Calcium:

In an aspect of the present invention, the composition comprises ioniccalcium.

The ionic calcium may be any calcium source suitable to be administratedto infants. For example, the ionic calcium may be one or more selectedfrom the group consisting of calcium citrate, calcium hydroxide, calciumoxide, calcium chloride, calcium carbonate, calcium gluconate, calciumphosphate, calcium diphosphate, calcium triphosphate, calciumglycerophosphate, calcium lactate and calcium sulphate.

In a preferred embodiment, the ionic calcium is one or more selectedfrom the group of calcium hydroxide, calcium chloride and calciumcitrate.

However, the ionic calcium may also be one or more from the group ofcalcium phosphate, calcium gluconate and calcium carbonate.

In the context of the present invention, ionic calcium refers to calciumwhich is not attached to proteins. Ionic calcium may also be known asfree calcium. Ionic calcium can be measured with a ion meter.

Calcium phosphate as such is not ionic, but a chemical reaction is madeunder digestion which releases some free calcium. For example calciumdiphosphate follows the following solubilituy equilibrium:

Ca₃(PO₄)₂(s)

3Ca²⁺(aq)+2PO₄ ³⁻(aq)

Thus, in the context of the present invention calcium phosphate isconsidered as a source of ionic calcium

In an embodiment of the invention, the composition comprises ioniccalcium in an amount of at least 1.7 mmol/L.

The inventors of the present invention has surprisingly found out thationic calcium in combination with a hydrolysed protein hydrolysate andan oil mix with palmitic acid provides an improved effect on calciumexcretion and thus may improve calcium absorption. Without being boundby any theory, the inventors of the present invention believes thationic calcium provides a significant calcium-peptide interaction withpeptides from hydrolysed protein and thus an indirect impact on thedifferent digestive enzyme. The calcium-peptides will cause a poorprotection of lipase as compared to intact proteins and to peptides notbound to calcium. Lipase is responsible for the release of sn-1(3)palmitic acids from triacylglycerols under digestion, and less sn-1(3)palmitic acids will be released when having a blend of hydrolysedprotein and calcium. This results in less formation of palmitic acidsoaps and thus more calcium which can be absorbed.

Further ionic calcium and magnesium are able to form bridges between thenegatively charged molecules of proteins and increase drastically theformation of heavy precipitates of whey proteins. These calcium andmagnesium bridges form much more easily during the heat denaturationwhich favours a more flexible open structure of the protein chains.

In a further embodiment of the invention, the composition comprisesionic calcium in an amount of at least 2.0 mmol/l, such as for exampleat least 2.2 mmol/L, preferably at least 2.5 mmol/L.

In a further embodiment of the present invention, the compositioncomprises ionic calcium in an amount of 1.7 to 5.0 mmol/L, such as 2.0to 4.5 mmol/L, preferably 2.5 to 4.2 mmol/L, such as 3.0 to 4.0 mmol/L.

Infant Formula:

In an aspect, the composition according to the invention is an infantformula.

However, in another aspect, the composition according to the inventionis a fortifier or supplement to human breast milk or to an infantformula, or the composition is a composition to be used in making up aninfant formula.

In the context of the present invention, the term “infant formula”refers to an infant formula comprising the nutrients normally requiredfor infants to obtain a suitable growth and wherein said infant formulacomprises proteins, carbohydrates, lipids, vitamins, minerals and traceelements.

The infant formula according to the present invention may be a starterformula for infants from the age of birth to 4 to 6 months and whichprovide complete nutrition for this age group (both for term and preterminfants). Further, the infant formula may be a follow-on formula forinfants between the ages of four to six months and twelve months whichare fed to the infants in combination with increasing amounts of thefoods, such as infant cereals and puréed fruits, vegetables and otherfoodstuffs as the process of weaning progresses.

The infant formula according to the present invention comprises a oilmix, wherein said oil mix comprises palmitic acid and wherein theproportion of sn-2 palmitic acid is at least 35% based on total palmiticacid. The amount of sn-1(3) palmitic acid is thus less than 35% byweight of total palmitic acid.

In an embodiment according to the invention, the oil mix comprises sn-2palmitate in an amount of 2-15% of total fatty acids present in the oilmix and sn-1(3) palmitate in an amount of 5-14% of total fatty acids inthe oil mix.

The oil mix may comprise other fatty acids than palmitic acid, mainlythese fatty acids are bound to triacylglycerols, but some may be presentas free fatty acids. Examples of other fatty acids present is stearicacid and myristic acid which like palmitic acid are saturated acids,monounsaturated fatty acids, such as oleic acid, and polyunsaturatedfatty acids, such as linoleic acid, alpha-linolenic acid, arachidonicacid, and docosahexaenoic acid.

In a preferred embodiment, the oil mix comprises stearic acids.

Typically, the infant formula comprises an oil mix in an amount of 20 to45 g per litre infant formula.

All the features and embodiments relating to the oil mix, palmitic acid,hydrolysed protein and ionic calcium described above in connection withthe composition according to the invention also applies to the infantformula.

Thus, in an embodiment, the infant formula comprises an oil mix, whereinthe oil mix comprises palmitic acid in an amount of 15 to 25% by weight,such as in an amount of 18 to 22% by weight.

The protein content in the infant formula (term and preterm) istypically between 1.6 to 3.6 g/100 kcal, but may be lower. The proteincontent in the starter formulas are in the lower range and the proteincontent of the follow-on formulas being in the upper end of the range.

In a further embodiment, the infant formula comprises proteins in anamount of 5.0 to 35.0 g per litre.

As mentioned earlier, the protein present in the infant formula may notall be hydrolysed protein, but a part of the protein in the infantformula may be another protein source which for example is intact.However, there must be a certain amount of hydrolysed protein present inthe infant formula to obtain the desired effect. Thus, the amount ofpartially hydrolysed proteins in the infant formula is at least 50% ofthe total amount of protein, such as at least 60% of the total amount ofprotein, preferably at least 70% of the total amount of protein, evenmore preferably at least 80% of the total protein. In an embodiment, theprotein present in the infant formula is essentially all partiallyhydrolysed protein.

By the term “essentially all” is meant that a major part, i.e.essentially all protein is partially hydrolysed, for example at least95% is partially hydrolysed, such as at least 98%.

The infant formula according to the present invention may also comprisea source of carbohydrates. The composition may comprise one or morecarbohydrate. The preferred source of carbohydrate is lactose althoughother carbohydrates such as saccharose, maltodextrin, and starch mayalso be added. Preferably, the carbohydrate present in the infantformula according to the present invention is between 9 and 14 g/100kcal. The carbohydrate present in the infant formula is preferablylactose.

The infant formula may also comprise all vitamins and mineralsunderstood to be essential in the daily diet in nutritionallysignificant amounts. Minimum requirements have been established forcertain vitamins and minerals. Examples of minerals, vitamins and othernutrients present in the nutritional composition include vitamin A,vitamin B1, vitamin B2, vitamin B6, vitamin E, vitamin K, vitamin C,vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid,choline, calcium, phosphor, iodine, iron, magnesium, copper, zinc,manganese, chloride, potassium, sodium, selenium, chromium, molybdenum,taurine, and L-carnitine. The minerals are usually added in the saltform.

Other components may be added to an infant formula, such as probiotics,prebiotics, emulsifiers and stabilizers.

Preparation:

The composition and infant formula according to the present inventionmay be prepared by any known or otherwise suitable manner. For example,an infant formula may be proposed by blending together a source ofprotein with a carbohydrate source and a lipid source in appropriateproportions. If used, emulsifiers may be included at this stage.Vitamins and minerals may be added at this stage, but may also be addedlater to avoid thermal degradation. Water, preferably water which hasbeen subjected to reverse osmosis or deionized water, may then be addedand mixed in to form a liquid mixture. The temperature of mixing ispreferably room temperature, but may also be higher. The liquid mixturemay then be thermally treated to reduce bacterial loads. The mixture maythen be homogenized.

If it is desired to produce a powdered composition, the homogenizedmixture is dried in a suitable drying apparatus, such as a spray drieror freeze drier and converted into powder.

Processes used in the manufacture of formulae for infants and youngchildren are based on the concept that the products must benutritionally adequate and microbiologically safe to consume. Thus,steps that eliminate or restrict microbiological growth are central toproduction processes. The processing technology for each specificformula is proprietary to the manufacturer but, in general, it involvesthe preservation of an oil-in-water (o/w) emulsion by dehydration in thecase of powder products or, sterilization in the case of ready-to-feedor concentrated liquid products. Powdered infant formula may be producedusing various processes, such as dry blending dehydrated ingredients toconstitute a uniform formula or hydrating and wet-mixing a mixture ofmacro-ingredients, such as fat, protein and carbohydrate ingredients andthen evaporating and spray drying the resultant mixture. A combinationof the two processes described above may be used where a base powder isfirst produced by wet-mixing and spray drying all or some of themacro-ingredients and then dry blending the remaining ingredients,including carbohydrate, minerals and vitamins and other micronutrients,to create a final formula. Liquid formulae are available in aready-to-feed format or as a concentrated liquid, which requiresdilution, normally 1:1, with water. The manufacturing processes used forthese products are similar to those used in the manufacture ofrecombined milk.

If it is desired to produce a liquid infant formula, the homogenizedmixture is filled into suitable containers, preferably aseptically.However, the liquid composition may also be retorted in the container,suitable apparatus for carrying out the filling and retorting of thisnature is commercially available.

Use of the Composition/Infant Formula:

The present invention is also directed to the composition or infantformula according to the present invention for use in administration toan infant or young child to reduce calcium excretion by said infant oryoung child. The composition or infant formulation is administered tothe infant by feeding the infant.

The infant could be any infant, i.e. both a term and preterm infant. Thecomposition or infant formula according to the invention mayadvantageously be administered to infants or young children. In aparticular embodiment, it is used for infants of less than 12 months,i.e. less than 6 months or less than 3 months. In one embodiment, thecomposition or infant formula is a preterm infant formula. In anotherembodiment, the composition or infant formula is designed forconsumption by infants from birth to 12 months.

The products of the invention can be in a fluid (liquid) form. These canbe sold ready to consume (without further dilution).

The products of the invention can be in the form of dehydrated powderswhich are prepared for consumption by reconstitution with water or milk.

The products of the invention can also be in the form of extruded snackproduct, especially intended for young children.

Further, the present invention is directed to the composition or infantformula according to the present invention for use in administration toan infant or young child to improve calcium homeostasis, increasecalcium absorption, increase calcium retention, increase calciumutilization and/or reduce the formation of insoluble palmitic acidsoaps.

Without being bound by theory, the increased calcium absorption isthought to have a beneficial physiological effect as well as an effecton bone mineralisation. The inventors of the present invention hassurprisingly found out that when using ionic calcium in combination withhydrolysed proteins and an oil mix comprising palmitic acid for use ininfant formulation, a synergistic effect is obtained and calcium is moreefficiently absorbed than when using a composition comprising an oil mixwith palmitic acid either with intact proteins or in combination withhydrolysed proteins. The hydrolysed proteins are without being bound byany theory believed to influence the effect of pancreatic lipase whichreleases palmitic acids from the sn-1 and sn-3 (alpha) position oftriacykglycerols. Thus, when hydrolysed proteins are present, lesssn-1(3) palmitic acid is released from the triacylglycerols and thusless free palmitic acids available to form soaps with calcium. Reducedpalmitic acid soaps may lead to reduced constipation and improvedgastrointestinal tolerance as compared to a standard infant formula.Hence, when less calcium is excreted, more calcium may be absorbed.

Further, ionic calcium in combination with hydrolysed proteins furtherdecreases the effect of lipase, since calcium forms bonds with peptidesfrom the hydrolysed proteins. Thus, hydrolysed proteins and ioniccalcium provides a synergistic effect such that less palmitic acid insn-1 and sn-3 position of triacylglycerols is released.

Thus, when hydrolysed proteins and ionic calcium are present, it willresult in less sn-2 palmitic acid released and thus less free palmiticacids released which can bind to free calcium. Therefore, less palmiticacid soaps are formed. Reduced palmitic acid soaps may lead to reducedconstipation and improved gastrointestinal tolerance as compared to astandard infant formula. Hence, when less calcium is excreted, morecalcium is absorbed.

Similar effect on calcium excretion can be obtained with a formulacomprising an oil mix comprising palmitic acid, hydrolysed proteins andionic calcium which is similar to the calcium excretion obtained withhuman milk.

The present invention is also directed to a composition or infantformulation according to the invention for use in administration to aninfant or young child to soften stools, prevent and/or reduce the riskof hard stools, digestive discomfort from hard stools, prevent and/orreduce the risk of constipation, decrease the frequency and duration ofcrying and fussing, and alleviate digestive discomfort and colic. Areduction in digestive discomfort can improve infant sleep duration,improve sleep quality and quantity, improve infant and parental qualityof life and reduce maternal anxiety.

Increased calcium absorption and retention can lead to increased bonemineralization. Increased calcium absorption is due to less formation ofcalcium soaps which are poorly absorbed, but on the contrary excreted.These palmitic acid soaps (calcium—fatty acid complexes) when excretedin the stool may cause the stool to be hard stool. Thus, an object ofthe present invention is also softening of stools.

Furthermore, an aspect of the present invention relates to thecomposition or infant formula according to the present invention for usein administration to an infant or young child to increase bonemineralization, increase bone strength, and increase bone mineraldensity.

Especially, for preterm infants or low or very low birth weight infantshave a high incidence of metabolic bone disease, 30-50%, and there aretherefore at need to increase bone mineralization in this type ofinfants. Metabolic bone disease increases the risk of fractures andgrowth failure. Inadequate calcium absorption is believed to be a mainfactor in metabolic bone disease.

It should be noted that embodiments and features described in thecontext of one of the aspects of the present invention also apply to theother aspects of the invention.

All patent and non-patent references cited in the present application,are hereby incorporated by reference in their entirety.

The invention will now be described in further details in the followingnon-limiting examples.

EXAMPLES

The following examples illustrates the specific embodiments of thecomposition and infant formula according to the invention and the use ofsaid composition and infant formula. The examples are given solely forthe purpose of illustration and are not to be construed as limitationsif the present invention, as many variations thereof are possiblewithout departing form the spirit of the invention.

Example 1

Example 1 describes a comparison of different studies conducted to studyinfants being fed with different infant formulas compared to a controlinfant formula

In group I, infants are fed with an infant formula comprising intactproteins and a fat blend comprising triacylglycerols with 35.9% ofpalmitic acid in the sn-2 position of triacylglycerol. The infants werefed with said infant formula for four weeks and then palmitate soaps(calcium-palmitate complexes), total fatty acid soaps and calcium in thestool were measured and compared to the amounts in a control infantformula.

In group II, infants, as in group I, are fed with an infant formulacomprising intact proteins and a fat blend comprising triacylglycerols,with 35.9% of palmitic acid in sn-2 position of triacylglycerol. Theinfants were fed with said infant formula for eight weeks and thenpalmitate soaps (calcium-palmitate complexes), total fatty acid soapsand calcium in the stool were measured and compared to the amounts in acontrol infant formula.

In group III, infants are fed with an infant formula comprisingpartially hydrolysed protein and a fat blend comprising triacylglycerolswith 35.9% of palmitic acid in sn-2 position of triacylglycerol. Theinfants were fed with said infant formula for eight weeks and thenpalmitate soaps (calcium-palmitate complexes), total fatty acid soapsand calcium in the stool were measured and compared to the amounts in acontrol infant formula.

The control infant formula comprises a fat blend having beta palmiticacid in an amount of about 11.7%. Further, the control formula comprisesa standard intact cow's milk protein as the protein source.

The calcium content in stool is also measured for a group of infants fedwith human milk.

The primary objective of the studies was to measure the amount ofcalcium and calcium-fatty acid soaps in the stool of infants fed withthe different infant formulas as compared to a control infant formula.

Both the three test formulas (in group I, II, and III) and the controlformula was prepared as a ready-to-feed liquid. The three test formulasand the control formula are described below in table 1A.

TABLE 1A Test formula Test formula Test formula Control in group I inGroup II in group III formula Amount 8.2 g/L 8.2 g/L 8.2 g/L 7.5 g/Lpalmitic acid 22.8% 22.8% 22.8% 21.3% in oil mix Amount sn-2 35.9% 35.9%35.9% 11.7% palmitate of total palmitic acid Amount sn-2 8.2% 8.2% 8.2%62.6% palmitate (% of total fat) Amount Sn- 14.6% 14.6% 14.6% 18.8% 1(3)palmitate (% of total fat)

The ionic calcium content in group III and the control group is shown inthe below table 1B:

Ionic calcium (mmol/L) Control 1.09 Test formula 3.65

The calcium excreted in the stools were measured for the three groupsand in the control formula.

Table 2 below shows the calcium stool content measured in the faeces ofinfants in group I, II and III. Further, table 2 shows the calcium stoolcontent in infants fed with the control and in infants fed with humanbreast milk. Table 2 furthermore shows the calcium content in thedifferent infant formulas

TABLE 2 Group I Group II Group III infant formula infant formula infantformula vs. control vs. control vs. control Stool ↑ 3.7% ↓ 5.2% ↓ 20.5%Calcium NS NS (p < 0.05) (% change) Stool Test Ctrl HM Test Ctrl HM TestCtrl ND calcium 39.4 38 17.6 30.7 32.4 20.3 25.6 32.3 (mean mg/g CalciumTest Ctrl HM Test Ctrl HM Test Ctrl ND content 42 42 NA 46 47 NA 46 47in test formula (mg/100 ml) NS = not significant; ND = not done; NA =not applicable

From table 2 it can be seen that the calcium content in the testformulas and control formulas are almost similar, i.e. no significantdifference.

Furthermore, table 2 shows that the infants fed the test formula ingroup III have a faecal calcium content more similar infants fed withhuman breast milk than infants fed test formula in group I and II. Thus,infants fed with an infant formula comprising moderate amounts of betapalmitic acid and partially hydrolysed proteins has a content of calciumexcreted in stools more similar to infants fed with human breast milk ascompared to infants fed with infant formulas with a high amount of betapalmitic acid and intact proteins.

Besides from the calcium content excreted in the stools, the fatty acidsoaps were also measured for the three groups. The content of calcium instools and the fatty acid soaps were measured and compared to thecontents in the control formula. The data can be seen in table 3.

TABLE 3 Group I Group II Group III infant formula infant formula infantformula vs. control vs. control vs. control Palmitate ↓ 15% ↓ 36% ↓ 41%soaps (p = 0.0028) (p < 0.0001)  (p < 0.0001) Total fatty  ↓ 8% ↓ 40% ↓28% acid soaps (NS) (p = 0.0001) (p < 0.01) Calcium in ↑ 3.7%  ↓ 5.2%  ↓20.5%  stool (NS) (NS) (p < 0.05) ↓: refers to a reduction as comparedto the control. ↑: refers to an increase as compared to the control.

The infants in the test groups are fed with the test infant formulas asthe sole nutrition.

From table 3, it is shown that the infant formula comprising acombination of beta palmitic acid (35.9%) and hydrolysed protein has animproved effect on reducing the amount of palmitic acid-calciumcomplexes (soaps) as compared to infants fed with an infant formulacomprising palmitic acid in an amount of 35.9 and intact proteins. Thus,it is shown that hydrolysed proteins have an effect on calcium excretionin stools when administered with beta palmitic acid in moderate amounts.The calcium excretion for the infants in group III was reduced by 20% ascompared to infants fed with the control infant formula, while there wasno significant difference in the calcium excretion for the infants ingroup I and II. Further, the faecal calcium content for the infants fedin group III was closer to the faecal calcium content in infants fedwith human breast milk.

1. A composition for use in formulas for infants or young childrencomprising partially hydrolysed protein, ionic calcium and an oil mixcomprising palmitic acid esterified to triacylglycerols, wherein thepalmitic acid comprises palmitic acid esterified in the sn-2 position oftriacylglycerols in an amount of at least 20% by weight of totalpalmitic acid.
 2. Composition according to claim 1, wherein the ioniccalcium is selected from the group consisting of calcium citrate,calcium hydroxide, calcium oxide, calcium chloride, calcium carbonate,calcium gluconate, calcium phosphate, calcium diphosphate, calciumtriphosphate, calcium glycerophosphate, calcium lactate, and calciumsulphate.
 3. Composition according to claim 1, wherein the compositioncomprises ionic calcium in an amount of at least 1.7 mmol/L. 4.Composition according to claim 1, wherein the partially hydrolysedprotein is partially hydrolysed whey protein.
 5. Composition accordingto claim 1, wherein the partially hydrolysed protein has a degree ofhydrolysation from 8% to 24%.
 6. Composition according to claim 1,wherein the oil mix comprises palmitic acid in an amount of at least 8%by weight of the total amount of fatty acids.
 7. Composition accordingto claim 1, wherein the ratio between the partially hydrolysed proteinand the sn-2 palmitic acid is from 1.5:1 to 35:1 based on weight. 8.(canceled)
 9. Composition according to claim 1, wherein the oil mixcomprises sn-2 palmitic acid in an amount of 15 to 25% by weight. 10.Composition according to claim 1, wherein the amount of partiallyhydrolysed proteins is at least 50% of the total amount of protein. 11.A method for use in administration to an infant or young child to reducecalcium excretion by the infant or young child comprising administeringa composition comprising partially hydrolysed protein, ionic calcium andan oil mix comprising palmitic acid esterified to triacylglycerols,wherein the palmitic acid comprises palmitic acid esterified in the sn-2position of triacylglycerols in an amount of at least 20% by weight oftotal palmitic acid.
 12. A method for use in administration to an infantor young child to increase improve calcium homeostasis, calciumabsorption, increase calcium retention, increase calcium utilizationand/or reduce the formation of palmitic acid soaps comprisingadministering a composition comprising partially hydrolysed protein,ionic calcium and an oil mix comprising palmitic acid esterified totriacylglycerols, wherein the palmitic acid comprises palmitic acidesterified in the sn-2 position of triacylglycerols in an amount of atleast 20% by weight of total palmitic acid. 13-15. (canceled)